Effect of VEGF-C siRNA and endostatin on ring formation and proliferation of esophageal squamous cell carcinoma lymphatic endothelial cells

Abstract

Yuping Zheng,1–3,* Miaomiao Sun,4,* Jinyan Chen,1,2 Lulu He,1,2 Na Zhao,1,2 Kuisheng Chen1,2 1Pathology Department, The First Affiliated Hospital of Zhengzhou University, 2Henan Key Laboratory of Tumor Pathology, 3Pathology Department, The Second Hospital of Shandong University, Jinan, 4Pathology Department, Henan Tumor Hospital, Zhengzhou, People’s Republic of China *These authors contributed equally to this work Objective: To study the effects of vascular endothelial growth factor C small interfering RNA and endostatin on esophageal squamous cell carcinoma-related ring formation in vitro and proliferation of lymphatic endothelial cells.Materials and methods: KYSE150 cells were subjected to analysis of cell transfection and endostatin operation. The groups were as follows: negative group, blank group, negative plus endostatin group, endostatin group, SG1 group, SG2 group, SG1 plus endostatin group, and SG2 plus endostatin group. The esophageal cancer-related microlymphatic endothelial cells were three-dimensionally cultured. Cell Counting Kit-8 (CCK-8) assay was employed to detect cell proliferation.Results: The negative group’s three-dimensional culture result was the highest, followed by the blank group, negative plus endostatin group, endostatin group, SG2 group, SG1 group, SG1 plus endostatin group, and SG2 plus endostatin group. The quantity of living cells in the blank group was the highest, followed by the negative control, endostatin, SG2, SG1, negative plus endostatin, SG1 plus endostatin, and SG2 plus endostatin groups. Conclusion: Both vascular endothelial growth factor C small interfering RNA and endostatin could inhibit ring formation in esophageal squamous cell carcinoma and proliferation of lymphatic endothelial cells. Keywords: esophageal squamous carcinoma cells, esophageal cancer-associated lymphatic endothelial cells, VEGF-C, ring formation, proliferation

Keywords

• Objective To investigate the effects of VEGF-C inRNA eukaryotic expression vector and expression of endostatin on esophageal squamous carcinoma-related ring formation in vitro and proliferation of lymphatic endothelial cells. Methods KYSE150 cells were subjected to analysis of cell transfection and endostatin operation. The conditional medium was prepared as well. Blank control group was for the conditional medium of esophageal cancer cells without transfection
• Negative control group was for the conditional medium of empty plasmid transfected esophageal cancer cells
• SG1 group was for the conditional medium of SG1 plasmid transfected with esophageal cancer cells
• SG2 group was for the conditional medium of SG2 plasmid transfected esophageal cancer cells
• Endostatin group was for the esophageal cancer cells conditional medium of optimum concentration endostatin processed
• There were also additional three groups
• of which were the negative plus endostatin group
• SG1 plus endostatin group and SG2 plus endostatin group. The esophageal cancer related microlymphatic endothelial cells were three-dimensional cultured and was calculated for the various cells formed tube-like structures. CCK-8 assay was employed to detect the cell proliferation condition. Result Generally comparison
• the three dimensional culture results of negative control group was the highest
• then followed by blank control group
• negative plus endostatin group
• endostatin group
• SG2 group
• SG1 group
• SG1 plus endostatin group
• SG2 plus endostatin group. The comparsion between SG1 plus endostatin group and SG2 endostatin group did not have statistical significance (P>0.05)
• the comparsion between every two of SG1 group
• SG2 group
• endostatin group
• negative plus endostatin group did not have statistical significance (P>0.05)
• the comparison between rest groups had statistical significance (P
• the alive cells quantity in blank control group were the largest
• then followed by negative control group
• endostatin group
• SG2 group
• SG1 group
• negative plus endostatin group
• SG1 plus endostatin group and SG2 plus endostatin group. There was no statistical significance between the comparison of SG1 group and SG2 group
• SG1 group and endostation group
• SG1 group and negative plus endostatin group
• SG2 group and endostation group
• SG2 group and negative plus endostatin group
• SG1 plus endostatin group and SG2 plus endostatin group (P >0.05). The comparison between rest groups had statistical significance (P